This core proposal is to exploit viral pathogen protein microarrays for the purpose of identifying novel hostvirus protein interacting partners that regulate virus-host species tropism, host innate immune response and mediate pathogenic zoonotic infections in man. We propose to utilize a newly developed technology, NAPPA (Nucleic Acid Programmable Protein Arrays), which is currently being developed at the Harvard Proteomic Facility for the human proteome. We are now completing the construction of the monkeypox virus (MPV) proteome array (strain Zaire), but will expand the repertoire of the microarrays to include other viral pathogens of interest to SERCEB and the RCE program: starting with, human coronaviruses, dengue and Chikungunya virus. The virus-specific protein microarrays will be used to screen for novel human-virus protein interactions, with emphasis initially on the human inflammasome pathway, the interferon (IFN) pathway, and tumor necrosis factor (TNF) signaling pathway members. Viral proteome microarrays also have utility for studying anti-viral immune responses, screening for novel viral drug targets, and developing novel diagnostic strategies. Through the use of this proteomic platform technology, we seek to address the hypothesis that there are specific protein-protein interactions between proteins from human viral pathogens and human antiviral elements that are critical for host species tropism and pathogenesis in primates and man. By identifying viral/host protein interactions that regulate tropism and disease virulence properties of these viruses, we seek to identify specific targets for the development of novel antiviral strategies, as well as to better define the regulators of viral-induced pathogenic syndromes, such as the virus-induced "cytokine storm" that is thought to underlie the pathogenesis associated with a variety of zoonotic viral diseases in man. This project is being pursued in collaboration with the Harvard Proteomics Facility (Director: Josh LaBaer), which is developing the NAPPA technology to create human and select bacterial pathogen protein microarrays.